Design and Direct Assembly of Synthesized Uracil-containing Non-clonal  DNA Fragments into Vectors by USERTM Cloning

Morten Jørgensen; Nikolai Wulff; Majse Nafisi; Deyang Xu; Cuiwei Wang; Sophie  Lambertz; Zeinu Belew; Hussam  Nour-Eldin

doi:10.21769/BioProtoc.2615

Bio-Protocol (Nov 2017)

Design and Direct Assembly of Synthesized Uracil-containing Non-clonal DNA Fragments into Vectors by USERTM Cloning

Morten Jørgensen,
Nikolai Wulff,
Majse Nafisi,
Deyang Xu,
Cuiwei Wang,
Sophie Lambertz,
Zeinu Belew,
Hussam Nour-Eldin

Affiliations

Morten Jørgensen: University of Würzburg, Institute for Molecular Plant Physiology and Biophysics, Würzburg, Germany
Nikolai Wulff: DynaMo Center, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Frederiksberg C, DenmarkCopenhagen Plant Science Center, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Frederiksberg C, Denmark
Majse Nafisi: Copenhagen Plant Science Center, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Frederiksberg C, DenmarkPlant Biochemistry Laboratory, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Frederiksberg C, Denmark
Deyang Xu: DynaMo Center, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Frederiksberg C, DenmarkCopenhagen Plant Science Center, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Frederiksberg C, Denmark
Cuiwei Wang: DynaMo Center, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Frederiksberg C, DenmarkCopenhagen Plant Science Center, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Frederiksberg C, Denmark
Sophie Lambertz: DynaMo Center, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Frederiksberg C, DenmarkCopenhagen Plant Science Center, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Frederiksberg C, Denmark
Zeinu Belew: DynaMo Center, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Frederiksberg C, DenmarkCopenhagen Plant Science Center, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Frederiksberg C, Denmark
Hussam Nour-Eldin: DynaMo Center, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Frederiksberg C, DenmarkCopenhagen Plant Science Center, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Frederiksberg C, Denmark

DOI: https://doi.org/10.21769/BioProtoc.2615
Journal volume & issue: Vol. 7, no. 22

Abstract

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This protocol describes how to order and directly assemble uracil-containing non-clonal DNA fragments by uracil excision based cloning (USER cloning). The protocol was generated with the goal of making synthesized non-clonal DNA fragments directly compatible with USERTM cloning. The protocol is highly efficient and would be compatible with uracil-containing non-clonal DNA fragments obtained from any synthesizing company. The protocol drastically reduces time and handling between receiving the synthesized DNA fragments and transforming with vector and DNA fragment(s).

Published in Bio-Protocol

ISSN: 2331-8325 (Online)
Publisher: Bio-protocol LLC
Country of publisher: United States
LCC subjects: Science: Biology (General)
Website: https://bio-protocol.org

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